Cell Free Screening Assay
OHSU # 1056
- Maureen Hoatlin, SM.Biochemistry & Molecular Biology, Medicine, MMG
- Alexandra Sobeck, SM.Biochemistry & Molecular Biology
- Stacie Stone, SM-Medicine
The Hoatlin Cell-Free Screening Assay is biochemical assay platform that was initially developed to study proteins in the Fanconi Anemia-Breast Cancer (FA-BRCA) pathway. Derangements in the FA-BRCA pathway are common in a number of cancers, including breast cancer, head and neck cancers, and ovarian cancer. This versatile assay can be used to screen for drugs that can either stimulate or inhibit the pathway. The assay is adaptable to high throughput screening.
The cell-free assay is based on the preparation of replicating extract from Xenopus eggs. The egg extract is naturally arrested in the exact part of the cell cycle when the FA proteins are known to function. The extract mimics the complex interactions of proteins that control DNA replication in an intact cell and is capable of supporting DNA repair and DNA replication under naturally-regulated control.
The FA proteins function together in a pathway, a group of proteins with interdependent functions, composed of a multi-protein core complex and downstream proteins. The coordinated activityof the proteins is required for function during the S-Phase, aperiod in which DNA is replicated, and after the DNA has been damaged. The interaction of the FA core complex and the downstream protein FANCD2 is believed to be a key step that functionally links the FA core proteins to cancer susceptibility proteins BRCA1 and BRCA2 (FANCD1) as well as other proteins.
|Published||Patent Cooperation Treaty||WO 2007/067261 A1|
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